From 2013.igem.org
Sep 27
Kojima
| Lane | DNA | Enzyme
|
| 1 | 1kbp ladder | --
|
| 2 | pSB1C3 | EcoRI+SpeI
|
| 3
|
| 4
|
| 6 | Pcon-attenuator-aptmer-DT | EcoRI+SpeI
|
| 7
|
| 8
|
| 10 | Pcon-attenuator-aptmer-DT | EcoRI+SpeI
|
| 11
|
| 12
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-attenuator-DT(EcoRI+SpeI) | 6.8 | 1.98 | 0.43
|
| pT181-attenuator(EcoRI+SpeI) | 7.5 | 1.84 | 0.06
|
| pSB1C3(EcoRI+SpeI) | 17.7 | 1.97 | 0.71
|
Electrophoresis
No name
| Lane | Sample | Enzyme1 | Enzyme2
|
| 1 | pSB4K5 | E | P
|
| 2 | pSB4K5 | - | -
|
| 3 | 1kbp ladder | - | -
|
| 4 | Pcon-attenuator | E | S
|
| 5 | Pcon-attenuator | - | -
|
File:Igku xxxxxx.xxx
| Lane | Sample | Enzyme1 | Enzyme2
|
| 1 | RBS-GFP-DT | X | P
|
| 2 | RBS-GFP-DT | - | -
|
| 3 | RBS-GFP-DT | E | X
|
| 4 | 1kbp ladder | - | -
|
| 5 | Ptet | E | S
|
| 6 | Ptet | - | -
|
File:Igku xxxxxx.xxx
Liquid Culture
No name
| Sample | medium
|
| Pcon-GFP | Amp
|
| RBS-GFP-DT | CP
|
| Pcon-attenuator | Amp
|
| Pcon-attenuator-aptamer-DT | Amp
|
| Pcon-tetR-DT | Amp
|
Ligation
No name
| state | Vector | Inserter | Ligation High ver.2
|
| experiment | 9/17 pSB1C3 (E+S) 10.0ng/µL | 10µL | 9/26 pT181 attenuator(E+S) 7.5ng/µL | 1.3µL | 3.5 µL
|
| experiment | 9/25 DT (E+X) 42.8ng/µL | 2.3µL | 9/22 Pcon-pT181 antisense(E+S) 46.7ng/µL | 1.5µL | 1.9 µL
|
| experiment | 9/27 pSB4K5(E+P) 12.9ng/µL | 7.8µL | 9/10 Pcon-attenuator(E+S) 13.8ng/µL | 8.0µL | 3.5 µL
|
| experiment | 9/27 pSB4K5(E+P) 12.9ng/µL | 7.8µL | 9/25 RBS-GFP-DT(X+P) 13.9ng/µL | 12µL | 3.5 µL
|
| experiment | 9/8 pSB4K5 (E+S) 17.6ng/µL | xx µL | 8/21 Pcon-GFP-DT(E+S) 11ng/µL | 12.6µL | xx µL
|
Liquid Culture
Hirano
| Sample | medium
|
| Pcon-spinach-DT | plusgrow Amp
|
| Pcon-tetRaptamer-DT | plusgrow Amp
|
| Pcon-antisense-spinach-DT | plusgrow Amp
|
| spinach-DT | plusgrow CP
|
Nakamoto
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| 9/26 Pcon-GFP-DT | 423.0 | 1.83 | 1.46
|
| 9/26 Pcon-GFP-DT K | 298.7 | 1.82 | 1.50
|
Electrophoresis
Miniprep
Tatsui
| DNA
|
| Pcon-aptamer-DT
|
| Pcon-spinach-DT
|
| Pcon-antisense
|
| Pcon-tetR-DT
|
| Pcon-attenuator-DT
|
| Pcon-antisense-spinach-DT
|
Transformation
Hirano
| Name1 | Name2 | Sample1(µL) | Sample2(µL) | Competent Cells(µL) | Total(µL)
|
| Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-attenuator-DT(1A2) | 2 | 1 | 30 | 33
|
| Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-antisense-spinach-DT(1A2) | 2 | 1 | 30 | 33
|
| Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-tetR aptamer-DT(1A2) | 2 | 1 | 30 | 33
|
| Pcon-RBS-GFP-DT(4K5) | Pcon-attenuator-DT(1A2) | 2 | 1 | 30 | 33
|
| Pcon-RBS-GFP-DT(4K5) | Pcon-antisense-spinach-DT(1A2) | 2 | 1 | 30 | 33
|
| Pcon-RBS-GFP-DT(4K5) | Pcon-tetR aptamer-DT(1A2) | 2 | 1 | 30 | 33
|
| Pcon-attenuator+RBS-GFP-DT(4K5) | -- | 2 | -- | 20 | 22
|
| Pcon-RBS-GFP-DT(4K5) | -- | 2 | -- | 20 | 22
|
| attenuator(1C3) | -- | 2 | -- | 20 | 22
|
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